Pea line 08270930

ABSTRACT

The invention provides seed and plants of the pea line designated 08270930. The invention thus relates to the plants, seeds and tissue cultures of pea line 08270930, and to methods for producing a pea plant produced by crossing a plant of pea line 08270930 with itself or with another pea plant, such as a plant of another line. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of a plant of pea line 08270930, including the seed, pod, and gametes of such plants.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of pea line 08270930.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include greateryield, resistance to insects or pests, tolerance to heat and drought,better agronomic quality, higher nutritional value, growth rate andfruit or pod properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different varieties produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines are developed byselfing and selection of desired phenotypes. The new lines are evaluatedto determine which of those have commercial potential.

Pea plants are able to reproduce by self-fertilization andcross-fertilization. Thus far, however, commercial pea varieties havebeen inbred lines prepared through self fertilization (McPhee, 2005).

Peas are one of the top vegetables used for processing in the UnitedStates; with approximately 90% of the grown pea acreage used forprocessed consumption (NASS Census of Agriculture 2002). The pea is anannual cool season plant, growing best in slightly acidic soil. Manycultivars reach maturity about 60 days after planting. Pea plants canhave both low-growing and vining cultivars. The vining cultivars growthin tendrils from the leaves of the plant, which coil around availablesupports. The pea pods form at the leaf axils of the plant.

As with other legumes, pea plants are able to obtain fixed nitrogencompounds from symbiotic soil bacteria. Pea plants therefore have asubstantially reduced fertilizer requirement compared to non-leguminouscrops. This advantage adds to their commercial value, particularly inview of increasing fertilizer costs, and has generated considerableinterest in the creation of new pea plant cultivars.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a pea plant of the linedesignated 08270930. Also provided are pea plants having all thephysiological and morphological characteristics of the pea linedesignated 08270930. Parts of the pea plant of the present invention arealso provided, for example, including pollen, an ovule, a seed, a pod,and a cell of the plant.

The invention also concerns the seed of pea line 08270930. The pea seedof the invention may be provided as an essentially homogeneouspopulation of pea seed of the line designated 08270930. Essentiallyhomogeneous populations of seed are generally free from substantialnumbers of other seed. Therefore, seed of line 08270930 may be definedas forming at least about 97% of the total seed, including at leastabout 98%, 99% or more of the seed. The population of pea seed may beparticularly defined as being essentially free from hybrid seed. Theseed population may be separately grown to provide an essentiallyhomogeneous population of pea plants designated 08270930.

In another aspect of the invention, a plant of pea line 08270930comprising an added heritable trait is provided. The heritable trait maycomprise a genetic locus that is, for example, a dominant or recessiveallele. In one embodiment of the invention, a plant of pea line 08270930is defined as comprising a single locus conversion. In specificembodiments of the invention, an added genetic locus confers one or moretraits such as, for example, herbicide tolerance, insect resistance,disease resistance, and modified carbohydrate metabolism. In furtherembodiments, the trait may be conferred by a naturally occurring geneintroduced into the genome of the line by backcrossing, a natural orinduced mutation, or a transgene introduced through genetictransformation techniques into the plant or a progenitor of any previousgeneration thereof. When introduced through transformation, a geneticlocus may comprise one or more genes integrated at a single chromosomallocation.

In another aspect of the invention, a tissue culture of regenerablecells of a pea plant of line 08270930 is provided. The tissue culturewill preferably be capable of regenerating pea plants capable ofexpressing all of the physiological and morphological characteristics ofthe line, and of regenerating plants having substantially the samegenotype as other plants of the line. Examples of some of thephysiological and morphological characteristics of the line 08270930include those traits set forth in the tables herein. The regenerablecells in such tissue cultures may be derived, for example, from embryos,meristems, cotyledons, pollen, leaves, anthers, roots, root tips,pistil, flower, seed and stalks. Still further, the present inventionprovides pea plants regenerated from a tissue culture of the invention,the plants having all the physiological and morphologicalcharacteristics of line 08270930.

In yet another aspect of the invention, processes are provided forproducing pea seeds, pods and plants, which processes generally comprisecrossing a first parent pea plant with a second parent pea plant,wherein at least one of the first or second parent pea plants is a plantof the line designated 08270930. These processes may be furtherexemplified as processes for preparing hybrid pea seed or plants,wherein a first pea plant is crossed with a second pea plant of adifferent, distinct line to provide a hybrid that has, as one of itsparents, the pea plant line 08270930. In these processes, crossing willresult in the production of seed. The seed production occurs regardlessof whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent pea plant, oftenin proximity so that pollination will occur for example, mediated byinsect vectors. Alternatively, pollen can be transferred manually. Wherethe plant is self-pollinated, pollination may occur without the need fordirect human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent pea plants into plants that bear flowers. A third step maycomprise preventing self-pollination of the plants, such as byemasculating the male portions of flowers, (i.e., treating ormanipulating the flowers to produce an emasculated parent pea plant).Self-incompatibility systems may also be used in some hybrid crops forthe same purpose. Self-incompatible plants still shed viable pollen andcan pollinate plants of other varieties but are incapable of pollinatingthemselves or other plants of the same line.

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent pea plants. Yet another step comprisesharvesting the seeds from at least one of the parent pea plants. Theharvested seed can be grown to produce a pea plant or hybrid pea plant.

The present invention also provides the pea seeds and plants produced bya process that comprises crossing a first parent pea plant with a secondparent pea plant, wherein at least one of the first or second parent peaplants is a plant of the line designated 08270930. In one embodiment ofthe invention, pea seed and plants produced by the process are firstgeneration (F₁) hybrid pea seed and plants produced by crossing a plantin accordance with the invention with another, distinct plant. Thepresent invention further contemplates plant parts of such an F₁ hybridpea plant, and methods of use thereof. Therefore, certain exemplaryembodiments of the invention provide an F₁ hybrid pea plant and seedthereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from line 08270930, the method comprising thesteps of: (a) preparing a progeny plant derived from line 08270930,wherein said preparing comprises crossing a plant of the line 08270930with a second plant; and (b) crossing the progeny plant with itself or asecond plant to produce a seed of a progeny plant of a subsequentgeneration. In further embodiments, the method may additionallycomprise: (c) growing a progeny plant of a subsequent generation fromsaid seed of a progeny plant of a subsequent generation and crossing theprogeny plant of a subsequent generation with itself or a second plant;and repeating the steps for an additional 3-10 generations to produce aplant derived from line 08270930. The plant derived from line 08270930may be an inbred line, and the aforementioned repeated crossing stepsmay be defined as comprising sufficient inbreeding to produce the inbredline. In the method, it may be desirable to select particular plantsresulting from step (c) for continued crossing according to steps (b)and (c). By selecting plants having one or more desirable traits, aplant derived from line 08270930 is obtained which possesses some of thedesirable traits of the line as well as potentially other selectedtraits.

In certain embodiments, the present invention provides a method ofproducing peas comprising: (a) obtaining a plant of pea line 08270930,wherein the plant has been cultivated to maturity, and (b) collectingpeas from the plant.

In still yet another aspect of the invention, the genetic complement ofthe pea plant line designated 08270930 is provided. The phrase “geneticcomplement” is used to refer to the aggregate of nucleotide sequences,the expression of which sequences defines the phenotype of, in thepresent case, a pea plant, or a cell or tissue of that plant. A geneticcomplement thus represents the genetic makeup of a cell, tissue orplant, and a hybrid genetic complement represents the genetic make up ofa hybrid cell, tissue or plant. The invention thus provides pea plantcells that have a genetic complement in accordance with the pea plantcells disclosed herein, and plants, seeds and plants containing suchcells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that line 08270930 could be identified by any of themany well known techniques such as, for example, Simple Sequence LengthPolymorphisms (SSLPs) (Williams et al., 1990), Randomly AmplifiedPolymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF),Sequence Characterized Amplified Regions (SCARs), Arbitrary PrimedPolymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by pea plant cells, tissues, plants,and seeds, formed by the combination of a haploid genetic complement ofa pea plant of the invention with a haploid genetic complement of asecond pea plant, preferably, another, distinct pea plant. In anotheraspect, the present invention provides a pea plant regenerated from atissue culture that comprises a hybrid genetic complement of thisinvention.

In still yet another aspect, the invention provides a plant of an inbredpea line that exhibits a combination of traits comprising long vines,normal foliage, two relatively straight pods per node, dark green podsand berries, resistance to races 1 and 2 of Fusarium wilt, fieldresistance to Ascochyta and good stress tolerance. In certainembodiments, the combination of traits may be defined as controlled bygenetic means for the expression of the combination of traits found inpea line 08270930.

In still yet another aspect, the invention provides a method ofdetermining the genotype of a plant of pea line 08270930 comprisingdetecting in the genome of the plant at least a first polymorphism. Themethod may, in certain embodiments, comprise detecting a plurality ofpolymorphisms in the genome of the plant. The method may furthercomprise storing the results of the step of detecting the plurality ofpolymorphisms on a computer readable medium. The invention furtherprovides a computer readable medium produced by such a method.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of error for the device or method being employed to determinethe value. The use of the term “or” in the claims is used to mean“and/or” unless explicitly indicated to refer to alternatives only orthe alternatives are mutually exclusive, although the disclosuresupports a definition that refers to only alternatives and to “and/or.”When used in conjunction with the word “comprising” or other openlanguage in the claims, the words “a” and “an” denote “one or more,”unless specifically noted. The terms “comprise,” “have” and “include”are open-ended linking verbs. Any forms or tenses of one or more ofthese verbs, such as “comprises,” “comprising,” “has,” “having,”“includes” and “including,” are also open-ended. For example, any methodthat “comprises,” “has” or “includes” one or more steps is not limitedto possessing only those one or more steps and also covers otherunlisted steps. Similarly, any plant that “comprises,” “has” or“includes” one or more traits is not limited to possessing only thoseone or more traits and covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of pea line 08270930. This line shows uniformityand stability within the limits of environmental influence for thetraits described hereinafter. Pea line 08270930 provides sufficient seedyield. By crossing with a distinct second plant, uniform F1 hybridprogeny can be obtained.

In specific embodiments, the invention provides plants that exhibit anumber of improved traits, such as long vines, normal foliage, tworelatively straight pods per node, dark green pods and berries,resistance to races 1 and 2 of Fusarium wilt, field resistance toAscochyta and good stress tolerance.

A. Origin and Breeding History of Pea Line 08270930

The crossing and selections by which pea line 08270930 was made can besummarized as follows:

Year 1 (Spring) P - cross made between parent varieties Year 1 (Fall)F1 - planted F1; allowed plants to self Year 2 (Spring) F2 - planted F2;selected individual plants Year 2 (Summer) F3 - planted F3; selectedindividual plants Year 3 (Spring) F4 - planted F4; selected individualplants Year 3 (Summer) F5 - planted F5; selected individual plants Year4 (Spring) F6 - planted F6; selected individual plants Year 5 (Spring)F7 - planted one line; observed that the line was uniform and stable;bulk collected seeds from this line; all subsequent seed increases arederived from this line Year 6 (Spring) F8 - space planted a stock of theselected F7 seeds; harvested progeny as 100 individual plant selectionsYear 7 (Spring) Planted 100 progeny rows for seed increase; observedthat the progeny were uniform and stable; progenies harvested in bulk

Selection criteria included characteristics related to productivity andquality and fit to market needs. Specific criteria included dark greenpods, good stress tolerance, resistance to powdery mildew, resistance toRaces 1 and 2 of the wilt fungus Fusarium oxysporum fsp pisi, resistanceto Ascochyta pisi, resistance to pea enation mosaic virus, good yieldpotential, uniform dark green frozen pod color and good harvest window.

Observations during years five and seven confirm that pea line 08270930is uniform and stable within commercially acceptable limits. As is truewith other snow pea varieties, a small percentage of off-types can occurwithin commercially acceptable limits for almost any characteristicduring the course of repeated multiplications. No variants are known tooccur.

B. Physiological and Morphological Characteristics of Pea Line 08270930

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of pea line 08270930. A description of the physiologicaland morphological characteristics of pea line 08270930 is presented inTable 1.

TABLE 1 Physiological and Morphological Characteristics of Line 08270930CHARACTERISTIC 08270930 1. Type edible-podded 2. Maturity node number offirst bloom 14 no. of days earlier than Wando  0 3. Plant Height high 85cm cm shorter than Oregon Sugar Pod II  0 4. Vine habit indeterminatebranching 1-2 branches (Little Marvel) internodes zig zag stockinessmedium (Thomas Laxton WR) number of nodes 20 5. Leaflets color darkgreen (Alderman) wax medium molding marbled (Alaska) number of leafletpairs three or more leaflet type normal 6. Stipules present clasping notmarbled size (compared with leaflets) larger color (compared withleaflets) same color dark green color chart value (RHS color chart) 137Asipule size large selected comparison variety Oregon Sugar Pod IIstipule size: medium color: dark green color chart value: 137A 7. FlowerColor venation greenish standard white wing white keel white 8. Podsshape slightly curved end blunt (Alaska) color dark green (Alderman)surface smooth, dull borne double length 8.3 cm width 18 mm no. seedsper pod  8 9. Seeds color dark green shape rounded surface smooth colorpattern monocolor primary color cream & green hilum color whitecotyledon color green grams per 100 seeds 36 10. Disease FusariumWilt-Race 1 resistant Powdery Mildew resistant Fusarium Wilt (NearWilt)-Race 2 resistant *These are typical values. Values may vary due toenvironment. Other values that are substantially equivalent are withinthe scope of the invention.C. Breeding Pea Line 08270930

One aspect of the current invention concerns methods for crossing thepea line with itself or a second plant and the seeds and plants producedby such methods. These methods can be used for propagation of line08270930, or can be used to produce hybrid pea seeds and the plantsgrown therefrom. Hybrid seeds are produced by crossing line 08270930with second pea parent line.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing line 08270930 followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with line08270930 and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny are heterozygous for locicontrolling the characteristic being transferred, but are like thesuperior parent for most or almost all other loci. The last backcrossgeneration would be selfed to give pure breeding progeny for the traitbeing transferred.

The line of the present invention is particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the line. In selecting a second plant to cross with08270930 for the purpose of developing novel pea lines, it willtypically be preferred to choose those plants which either themselvesexhibit one or more selected desirable characteristics or which exhibitthe desired characteristic(s) when in hybrid combination. Examples ofpotentially desirable traits include, but are not necessarily limitedto, improved resistance to viral, fungal, and bacterial pathogens,improved insect resistance, pod morphology, herbicide tolerance,environmental tolerance (e.g. tolerance to temperature, drought, andsoil conditions, such as acidity, alkalinity, and salinity), growthcharacteristics, nutritional content, taste, and texture. Improved tasteand texture applies not only to the peas themselves, but also to thepods of those varieties yielding edible pods. In peas, as in otherlegumes, taste and nutritional content are particularly affected by thesucrose and starch content.

Among examples of fungal diseases of particular concern in peas areAscochyta pisi, Cladosporium pisicola (leaf spot or scab), Erysiphepolygoni (powdery mildew), Fusarium oxysporum (wilt), Fusarium solani(Fusarium root rot), Mycosphaerella pinodes (Mycospharella blight),Peronospora viciae (downy mildew), Phythium sp. (pre emergencedamping-off), Botrytis cinerea (grey mold), Aphanomyces euteiches(common root rot), Thielaviopsis basicola (black root rot), andSclerotina sclerotiorum (sclerotina white mold). Pea plant viraldiseases include: Bean yellow mosaic virus (BYMV), Bean leaf roll virus(BLRV), Pea Early Browning Virus (PEBV), Pea Enation Mosaic virus(PEMV), Pea Mosaic Virus (PMV), Pea seed-borne Mosaic Virus (PSbMV) andPea Streak Virus (PSV). An important bacterial disease affecting peaplants is caused by Pseudomonas pisi (bacterial blight), (Muehlbauer etal., 1983; Davies et al. 1985; van Emden et al., 1988).

Insect pests that may be of particular concern in peas include, forexample, Aphis cracivora (Groundnut aphid), Acyrthosiphon pisum (Peaaphid), Kakothrips robustus (Pea thrips), Bruchis pisorum (Pea seedbeetle), Callosobruchus chinensis (Adzuki bean seed beetle), Apion sp.(Seed weevil), Sitona lineatus (Bean weevil), Contarina pisi (Peamidge), Helicoverpa armigera (African bollworm), Diachrysia obliqua (Podborer), Agriotis sp. (Cut worms), Cydia nigricana (Pea moth), Phytomuzahorticola (Leaf minor), Heliothis Zea (American bollworm), EtiellaZinckenella (Lima bean pod borer), Ophiomyia phaseoli (Bean fly), Deliaplatura (Bean seed fly), Tetranychus sp. (Spider mites), Pratylenchuspenetrants (Root lesion nematodes), Ditylenchus dipsaci (Stem nematode),Heterodera goettingiana (Pea cyst nematode), and Meloidogyne javanica(Root knot nematode), (van Emden et al., 1988; Muehlbauer et al., 1983).

D. Performance Characteristics

The performance characteristics of the line 08270930 were the subject ofan objective analysis of the performance traits of the line relative toother lines. The results of the analysis are presented below.

TABLE 2 Analysis of Performance Characteristics For Line 08270930 PlantPods Plant Height Pod per Viable Ease yield/ Variety Location Habit (cm)Lodging Set node Pods of Harvest Maturity Rating Taste Color Texture Haoverall SV08270930 Kenya 5 100 1 1 3 3 1 67 1 1 3 3 33 1 SV08790490Kenya 5 120 1 1 3 7 1 67 5 1 5 1 28 5E. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those pea plants which are developed by a plantbreeding technique called backcrossing, wherein essentially all of thedesired morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parental peaplant which contributes the locus for the desired characteristic istermed the nonrecurrent or donor parent. This terminology refers to thefact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental pea plant to whichthe locus or loci from the nonrecurrent parent are transferred is knownas the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a pea plant isobtained wherein essentially all of the desired morphological andphysiological characteristics of the recurrent parent are recovered inthe converted plant, in addition to the single transferred locus fromthe nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny pea plants of a backcross in which 08270930is the recurrent parent comprise (i) the desired trait from thenon-recurrent parent and (ii) all of the physiological and morphologicalcharacteristics of pea line 08270930 as determined at the 5%significance level when grown in the same environmental conditions.

Pea varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,male sterility, herbicide resistance, resistance to bacterial, fungal,or viral disease, insect resistance, restoration of male fertility,modified fatty acid or carbohydrate metabolism, and enhanced nutritionalquality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. An example of a dominant trait is the downy mildewresistance trait. For this selection process, the progeny of the initialcross are sprayed with downy mildew spores prior to the backcrossing.The spraying eliminates any plants which do not have the desired downymildew resistance characteristic, and only those plants which have thedowny mildew resistance gene are used in the subsequent backcross. Thisprocess is then repeated for all additional backcross generations.

Selection of pea plants for breeding is not necessarily dependent on thephenotype of a plant and instead can be based on genetic investigations.For example, one can utilize a suitable genetic marker which is closelygenetically linked to a trait of interest. One of these markers can beused to identify the presence or absence of a trait in the offspring ofa particular cross, and can be used in selection of progeny forcontinued breeding. This technique is commonly referred to as markerassisted selection. Any other type of genetic marker or other assaywhich is able to identify the relative presence or absence of a trait ofinterest in a plant can also be useful for breeding purposes. Proceduresfor marker assisted selection applicable to the breeding of pea are wellknown in the art. Such methods will be of particular utility in the caseof recessive traits and variable phenotypes, or where conventionalassays may be more expensive, time consuming or otherwisedisadvantageous. Types of genetic markers which could be used inaccordance with the invention include, but are not necessarily limitedto, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al.,1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., 1998).

F. Plants Derived from Pea Line 08270930 by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into the pea line of the invention ormay, alternatively, be used for the preparation of transgenes which canbe introduced by backcrossing. Methods for the transformation of plants,including pea plants, are well known to those of skill in the art (see,e.g., Schroeder et al., 1993). Techniques which may be employed for thegenetic transformation of pea plants include, but are not limited to,electroporation, microprojectile bombardment, Agrobacterium-mediatedtransformation and direct DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

A particularly efficient method for delivering transforming DNA segmentsto plant cells is microprojectile bombardment. In this method, particlesare coated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species. An illustrativeembodiment of a method for delivering DNA into plant cells bymicroprojectile bombardment is the Biolistics Particle Delivery System,which can be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target pea cells. The screen disperses the particles sothat they are not delivered to the recipient cells in large aggregates.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., 1985). Moreover, recent technological advances in vectorsfor Agrobacterium-mediated gene transfer have improved the arrangementof genes and restriction sites in the vectors to facilitate theconstruction of vectors capable of expressing various polypeptide codinggenes. The vectors described have convenient multi-linker regionsflanked by a promoter and a polyadenylation site for direct expressionof inserted polypeptide coding genes. Additionally, Agrobacteriumcontaining both armed and disarmed Ti genes can be used fortransformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055).Agrobacterium-mediated transformation is a particularly beneficialmethod for producing recombinant pea-plants. Transformed pea plants maybe obtained by incubating pea explant material with Agrobacteriumcontaining the DNA sequence of interest (U.S. Pat. No. 5,286,635; U.S.Pat. No. 5,773,693).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986;Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plantsand expression of foreign genetic elements is exemplified in Choi et al.(1994), and Ellul et al. (2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for pea plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., 1985), including monocots(see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); atandemly duplicated version of the CaMV 35S promoter, the enhanced 35Spromoter (P-e35S) the nopaline synthase promoter (An et al., 1988), theoctopine synthase promoter (Fromm et al., 1989); and the figwort mosaicvirus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and anenhanced version of the FMV promoter (P-eFMV) where the promotersequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus19S promoter, a sugarcane bacilliform virus promoter, a commelina yellowmottle virus promoter, and other plant DNA virus promoters known toexpress in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can beused for expression of an operably linked gene in plant cells, includingpromoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g.,pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter,Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter,Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte etal., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., 1987; Schernthaner et al., 1988; Bustos et al., 1989).

Exemplary nucleic acids which may be introduced to the pea lines of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a pea plant according to the invention.Non-limiting examples of particular genes and corresponding phenotypesone may choose to introduce into a pea plant include one or more genesfor insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pesttolerance such as genes for fungal disease control, herbicide tolerancesuch as genes conferring glyphosate tolerance, and genes for qualityimprovements such as yield, nutritional enhancements, environmental orstress tolerances, or any desirable changes in plant physiology, growth,development, morphology or plant product(s). For example, structuralgenes would include any gene that confers insect tolerance including butnot limited to a Bacillus insect control protein gene as described in WO99/31248, herein incorporated by reference in its entirety, U.S. Pat.No. 5,689,052, herein incorporated by reference in its entirety, U.S.Pat. Nos. 5,500,365 and 5,880275, herein incorporated by reference ittheir entirety. In another embodiment, the structural gene can confertolerance to the herbicide glyphosate as conferred by genes including,but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPSgene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, hereinincorporated by reference in its entirety, or glyphosate oxidoreductasegene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporatedby reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., 1991). The RNA could also be a catalytic RNA molecule (i.e., aribozyme) engineered to cleave a desired endogenous mRNA product (seefor example, Gibson and Shillito, 1997). Thus, any gene which produces aprotein or mRNA which expresses a phenotype or morphology change ofinterest is useful for the practice of the present invention.

G. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing wherein essentially allof the morphological and physiological characteristics of an inbred arerecovered in addition to the characteristics conferred by the singlelocus transferred into the inbred via the backcrossing technique. By“essentially all,” it is meant that all of the characteristics of aplant are recovered that are otherwise present when compared in the sameenvironment and save for the converted locus, other than an occasionalvariant trait that might arise during backcrossing or directintroduction of a transgene. A single locus may comprise one gene, or inthe case of transgenic plants, one or more transgenes integrated intothe host genome at a single site (locus).

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a pea plant by transformation.

H. Deposit Information

A deposit of pea line 08270930, disclosed above and recited in theclaims, has been made with the American Type Culture Collection (ATCC),10801 University Blvd., Manassas, Va. 20110-2209. The date of depositwas Feb. 24, 2009. The accession number for those deposited seeds of pealine 08270930 is ATCC Accession No. PTA-9848. Upon issuance of a patent,all restrictions upon the deposit will be removed, and the deposit isintended to meet all of the requirements of 37 C.F.R. §1.801-1.809. Thedeposit will be maintained in the depository for a period of 30 years,or 5 years after the last request, or for the effective life of thepatent, whichever is longer, and will be replaced if necessary duringthat period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference:

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1. A seed of pea line 08270930, a sample of seed of said line havingbeen deposited under ATCC Accession Number PTA-9848.
 2. A plant of pealine 08270930, a sample of seed of said line having been deposited underATCC Accession Number PTA-9848.
 3. A plant part of the plant of claim 2.4. The plant part of claim 3, wherein said part is selected from thegroup consisting of a pod, pollen, an ovule and a cell.
 5. A pea plant,or a part thereof, having all the physiological and morphologicalcharacteristics of the pea plant of claim
 2. 6. A tissue culture ofregenerable cells of pea line 08270930, a sample of seed of said linehaving been deposited under ATCC Accession Number PTA-9848.
 7. Thetissue culture according to claim 6, comprising cells or protoplastsfrom a plant part selected from the group consisting of embryos,meristems, cotyledons, pollen, leaves, anthers, roots, root tips,pistil, flower, seed and stalks.
 8. A pea plant regenerated from thetissue culture of claim 6, wherein the regenerated plant expresses allof the physiological and morphological characteristics of pea line08270930, a sample of seed of said line having been deposited under ATCCAccession Number PTA-9848.
 9. A method of producing seed, comprisingcrossing the plant of claim 2 with itself or a second plant.
 10. Themethod of claim 9, wherein the plant of pea line 08270930 is the femaleparent.
 11. The method of claim 9, wherein the plant of pea line08270930 is the male parent.
 12. An Fl hybrid seed produced by themethod of claim
 9. 13. An Fl hybrid plant produced by growing the seedof claim
 12. 14. A method for producing a seed of a line08270930-derived pea plant comprising the steps of: (a) crossing a peaplant of line 08270930 with a second pea plant, a sample of seed of saidline having been deposited under ATCC Accession Number PTA-9848; and (b)allowing seed of a 08270930-derived pea plant to form.
 15. The method ofclaim 14, further comprising the steps of: (c) crossing a plant grownfrom said 08270930-derived pea seed with itself or a second pea plant toyield additional 08270930-derived pea seed; (d) growing said additional08270930-derived pea seed of step (c) to yield additional08270930-derived pea plants; and (e) repeating the crossing and growingsteps of (c) and (d) to generate further 08270930-derived pea plants.16. A method of vegetatively propagating a plant of pea line 08270930comprising the steps of: (a) collecting tissue capable of beingpropagated from a plant of pea line 08270930, a sample of seed of saidline having been deposited under ATCC Accession Number PTA-9848; (b)cultivating said tissue to obtain proliferated shoots; and (c) rootingsaid proliferated shoots to obtain rooted plantlets.
 17. The method ofclaim 16, further comprising growing plants from said rooted plantlets.18. A method of introducing a desired trait into pea line 08270930comprising: (a) crossing a plant of line 08270930 with a second peaplant that comprises a desired trait to produce Fl progeny, a sample ofseed of said line 08270930 having been deposited under ATCC AccessionNumber PTA- 9848; (b) selecting an Fl progeny that comprises the desiredtrait; (c) crossing the selected F1 progeny with a plant of line08270930 to produce backcross progeny; (d) selecting backcross progenycomprising the desired trait and the physiological and morphologicalcharacteristic of pea line 08270930; and (e) repeating steps (c) and (d)three or more times to produce selected fourth or higher backcrossprogeny that comprise the desired trait and essentially all of thephysiological and morphological characteristics of pea line 08270930when grown in the same environmental conditions.
 19. A pea plantproduced by the method of claim
 18. 20. A method of producing a plant ofpea line 08270930 comprising an added desired trait, the methodcomprising introducing a transgene conferring the desired trait into aplant of pea line 08270930, a sample of seed of said line 08270930having been deposited under ATCC Accession Number PTA-9848.
 21. Aprogeny plant of the plant of claim 2 that comprises all of thephysiological and morphological characteristics of pea line 08270930, asample of seed of said line having been deposited under ATCC AccessionNumber PTA-9848.
 22. A seed that produces the plant of claim
 21. 23. Amethod of producing peas comprising: (a) obtaining the plant of claim 2,wherein the plant has been cultivated to maturity, and (b) collectingpeas from the plant.